title: Analysis of carboxy terminal domain of metalloprotease of elastolytic Aeromonas hydrophila.
abstract: We examined the ability of Aeromonas hydrophila to lyse elastin. Eight of 13 strains showed elastolytic activity on agar medium containing elastin and 5 strains did not. In order to examine the involvement of the metalloprotease of A. hydrophila (AMP) in elastolytic activity, we made the amp-deletion mutant strain from an elastolytic strain. The elastolytic activity of the strain decreased with this deletion. The analysis of AMP released into the culture supernatant showed that AMP appeared outside of the cell as the intermediate consisting of a mature domain and carboxy terminal (C-terminal) propeptide domain. Further analysis showed that the intermediate has the ability to lyse elastin and that loss of the C-terminal domain causes loss of the elastolytic activity of the intermediate. We then determined the nucleotide sequence of the amps of all strains used in this study. Phylogenetic analysis revealed that these AMPs were divided into three groups. The AMPs from elastolytic strains belong to group I or group II, and AMPs from non-elastolytic strains belong to group III. The distance between group I and group II is small, but group III is located separately from groups I and II. Comparison of the amino acid residues of the C-terminal domain revealed that there are 13 amino acid residues specific to the C-terminal domain of group III. This indicates that the conformation of the C-terminal propeptide domain formed by these specific amino acid residues is important for AMP to express elastolytic activity.
authors.affiliation: Laboratory of Protein Function, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700–8530, Japan. takahashi@pharm.okayama-u.ac.jp
authors.forename: Eizo
authors.initials: E
authors.lastname: Takahashi
authors.forename: Hidetomo
authors.initials: H
authors.lastname: Kobayashi
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Yamanaka
authors.forename: Mayu
authors.initials: M
authors.lastname: Nakanishi
authors.forename: Arisa
authors.initials: A
authors.lastname: Tateishi
authors.forename: Takemi
authors.initials: T
authors.lastname: Abe
authors.forename: Sakae
authors.initials: S
authors.lastname: Arimoto
authors.forename: Tomoe
authors.initials: T
authors.lastname: Negishi
authors.forename: Keinosuke
authors.initials: K
authors.lastname: Okamoto
mesh_terms: D016980:Aeromonas hydrophila / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D002364:Caseins / Q000378:metabolism; D020134:Catalytic Domain; D004549:Elastin / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D045726:Metalloproteases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D011487:Protein Conformation; D016415:Sequence Alignment
chemical_list: D002364:Caseins; D004549:Elastin; D045726:Metalloproteases